JP4620652B2 - Novel polygalatenoside compound and antidepressant containing the same - Google Patents

Description

  The present invention relates to a novel compound purified from an aqueous extract of Polygala and to the use of the compound as an antidepressant.

  Mental disorders such as anxiety and neurosis have increased in recent years, and various antidepressants are currently used. Recently developed antidepressants, such as venlafaxine (a norepinephrine reuptake inhibitor) and bupropion (a norepinephrine / dopamine reuptake inhibitor), interact with multiple receptors in the central nervous system. This has been demonstrated by numerous studies.

  Yuan Zhi (Roots of the Department of Surgeonidae, Polygala tenuifolia Wild) has therapeutic effects such as sedation, antipsychotic action, cognitive improvement, neuroprotection and anti-inflammatory action on the central nervous system. Is an important herbal prescription. Long-distance is also applied to insomnia, nervous breakdown, amnesia, palpitations associated with anxiety, emotional instability, disorientation, and also used to prevent dementia and forgetfulness. Various xanthones, saponins, and oligosaccharide esters extracted from distant places have been reported (for example, Non-Patent Documents 1 to 9).

  Patent Document 1 discloses an antidepressant composition comprising a therapeutically effective amount of an active ingredient as a mixture of a pharmaceutically acceptable active ingredient with a carrier or diluent. The active ingredient is: i) a polar solvent extract of sunflower in which the polar solvent is water or a mixture of water and methanol or ethanol; ii) an aqueous fraction extracted from a polar solvent extract extracted with an organic solvent; iii) an organic eluate obtained by using a polar solvent extract or aqueous fraction in a reverse phase chromatographic column and then eluting the column with water and organic solvent, or iv) less than 30000 daltons in the organic eluate A filtrate having a molecular weight.

In Patent Document 2, for treating a human suffering from a physical or mental disease, disorder, or condition in which it is effective to inhibit the reuptake of norepinephrine, or for preventing suffering from such a disease Methods and compositions are disclosed. The above physical or mental illnesses, disorders, and conditions are addictive disorders (including those caused by alcohol, nicotine, and other psychoactive substances) and withdrawal syndromes, adjustment disorders (depressed feelings, anxiety, concurrent anxiety and feeling of depression, behavior Disabilities, behavioral and emotional disorders), age-related learning and mental disorders (including Alzheimer's disease), anorexia, lethargy, general attention to lack of attention (or other cognitive) disorders, attention Deficit hyperactivity disorder (ADHD), bipolar disorder, polyphagia, chronic fatigue syndrome, chronic or acute stress, chronic pain, behavioral disorder, mood circulation disorder, depression (adolescent depression, minor depression Including), dysthymic disorders, fibromyalgia and other somatic disorders (incorporation disorders, conversion disorders, pain disorders, psychosis, body dysmorphic disorders, indistinguishable body expression disorders, and unspecified Body expression disorder) Generalized anxiety disorder (GAD), incontinence (ie stress incontinence, stress incontinence, mixed cause incontinence), inhalation disorder, addiction (alcoholism), mania, migraine, obesity (ie obesity or excessiveness) Weight loss patients), obsessive compulsive disorder and related spectrum disorder, rebellious challenge disorder, panic disorder, peripheral neuropathy, posttraumatic stress disorder, premenstrual dysphoric disorder (ie premenstrual syndrome and Late luteal phase dysphoria), psychiatric disorders (including schizophrenia, schizophrenic emotional disorder and schizophrenia-like disorder), seasonal affective disorders, sleep disorders (such as narcosis, nocturnal urine disease), social phobia (social anxiety) Disorders), specific developmental disorders, selective serotonin reuptake inhibitor (SSRI) resistant syndrome (ie, initially effective, but then The effect of SRI therapy will not last patient), and tic disorders (e.g., selected from the group consisting of Tourette's syndrome).
UK Patent Application Publication No. 2383951 US Pat. No. 6,642,235 Fujita, T .; Liu, D .; Y. Ueda, S .; Takeda, Y .; , Phytochemistry 1992, 31, 3997-4000. Ikeya, Y .; Sugama, K .; Okada, M .; Mitsuhashi, H .; , Phytochemistry 1991, 30, 2061-2065 Ikeya, Y .; Sugama, K .; Okada, M .; Mitsuhashi, H .; , Chem. Pharm. Bull. 1991, 39, 2600-2605 Miyase, T .; Iwata, Y .; Ueno, A .; , Chem. Pharm. Bull. 1991, 39, 3082-3084 Jiang, Y. et al. Tu, P .; F. , Phytochemistry 2002, 60, 813-816 Sakuma, S .; Shojji, J .; , Chem. Pharm. Bull. 1981, 30, 810-821 Jiang, Y. et al. Tu, P .; , Chem. Pharm. Bull. 2005, 53, 1164-1166 Jiang, Y. et al. Tu, P .; J. et al. , Asian Nat. Prod. Res. 2003, 5, 279-283 Jiang, Y. et al. Zhang, W .; Tu, P .; Xu, X. J. et al. Nat. Prod. 2005, 68, 875-879

  The first object of the present invention is to provide a novel compound useful as an active ingredient of an antidepressant.

  The second object of the present invention is to provide the use of the novel compounds of the present invention as an antidepressant.

In the present invention, [ ¹²⁵ I] RTI-55 (3β- (4-iodophenyl) -tropan-2β-methyl carboxylate was used as the inhibitory action of methanol eluate of the water-soluble extract of P. tenufolia root. The binding of the ester) to the membrane protein was examined. The inventors of the present application isolated five novel oligosaccharide derivatives (compounds 1 to 5) from an extract of P. tenuifolia by bioassay, and these were converted to a norepinephrine transporter (hereinafter referred to as NET) [ ¹²⁵ I] RTI-55 binding was found to be inhibited. Among the compounds, the compounds 1 and 2 were found to be more effective than the other compounds.

  The novel compound extracted from the citrus tree of the present invention is useful as an active ingredient of an antidepressant.

  The present invention provides a compound represented by the following chemical formula (1) or a pharmaceutically acceptable salt. In the present specification, the compound represented by the following chemical formula (1) or a pharmaceutically acceptable salt may be collectively referred to as “polygalatenosides”.

In the chemical formula (1), R, R ′ and R ″ are each independently a hydrogen atom or R ₁ (in this case, R, R ′ and R ″ are not all hydrogen atoms), and R ₁ Is the following chemical formula (2);

In the formula, R ₂ represents one selected from the group consisting of a hydrogen atom, a C1-6 alkyl group, a C1-6 alkoxy group, or a halogen atom. Examples of C1-6 alkyl groups include methyl, ethyl, propyl, isopropyl, n-butyl, pentyl, neopentyl, sec-butyl, tert-butyl, 1,2-dimethylpropyl, Examples thereof include an n-hexyl group, a cyclo-hexyl group, a 1,3-dimethylbutyl group, a 1-iso-propylpropyl group, and a 1,2-dimethylbutyl group. C1-6 alkoxy groups include methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, pentoxy, neopentoxy, sec-butoxy, tert-butoxy, 1,2-dimethylpropoxy N-hexyloxy group, cyclo-hexyloxy group, 1,3-dimethylbutoxy group, 1-iso-propylpropoxy group, 1,2-dimethylbutoxy group and the like. Examples of the halogen atom include a fluorine atom, a chlorine atom, a bromine atom and an iodine atom.

Preferably R ₂ is a hydrogen atom.

Preferably, any one of R, R ′ and R ″ is R ₁ and the other two are hydrogen atoms. That is, preferably R is R ₁ and R ′ and R ″ are hydrogen atoms. is there. Preferably, R ′ is R ₁ and R and R ″ are hydrogen atoms. Preferably, R ″ is R ₁ and R and R ′ are hydrogen atoms.

  The polygalatenoside of formula (1) includes a racemic mixture of optically active compounds or optically pure R and S stereoisomers.

  The present invention further comprises an antidepressant composition comprising a therapeutically effective amount of the polygalatenoside represented by the above formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient together with a pharmaceutically acceptable carrier and diluent. provide.

  The antidepressant composition of the present invention can be administered to patients orally or parenterally as a composition formulated with a pharmaceutically acceptable carrier or dissolved or suspended in a pharmaceutically acceptable diluent.

  When the agent is used for oral administration, the polygalatenoside is used as an appropriate additive, for example, lactose, sucrose, mannitol, corn starch, synthetic or natural gum, excipients such as crystalline cellulose, starch, cellulose derivatives. Binders such as gum arabic, gelatin, polyvinyl pyrrolidone, disintegrants such as carboxymethyl cellulose cellulose, sodium carboxymethyl cellulose, starch, corn starch, sodium alginate, and lubricants such as talc, magnesium stearate, sodium stearate A solid preparation such as tablets, powders (powder), pills, and granules can be prepared by mixing with fillers or diluents such as calcium carbonate, sodium carbonate, calcium phosphate, and sodium phosphate. it can. Moreover, it is good also as a capsule using a hard or soft gelatin capsule. These solid preparations may be enteric-coated using a coating base such as hydroxypropylmethylcellulose phthalate, hydroxypropylmethylcellulose acetate succinate, cellulose acetate phthalate, and methacrylate copolymer. Further, the polygalatenoside is dissolved in a generally used inert diluent such as purified water, and if necessary, this solution is infiltrated, emulsifier, dispersion aid, surfactant, sweetener, flavor, Liquid preparations such as syrups and elixirs can be obtained by appropriately adding aromatic substances.

  When the antidepressant composition of the present invention is used for parenteral administration, the polygalatenoside is used in purified water, an appropriate buffer such as a phosphate buffer, physiological saline such as physiological saline, Ringer's solution, and lock solution. Sterile aqueous solutions, non-aqueous solutions, suspensions, liposomes or emulsions suitably combined with physiological salt solutions, ethanol, glycerin and commonly used surfactants, preferably as sterile injectable solutions for injection or spraying, It is administered intravenously, subcutaneously, intramuscularly, intraperitoneally, enterally, intrabronchially and the like. In this case, the liquid preparation preferably has a physiological pH, preferably a pH in the range of 6-8. Furthermore, the antidepressant composition of the present invention can be administered as a suppository using a pellet or an suppository base.

  Among the above-mentioned, a preferable formulation, dosage form, etc. are selected by the doctor in charge.

  The dose of the antidepressant composition of the present invention varies depending on the age, weight and symptoms of the patient, the intended dosage form and method, therapeutic effect, treatment period, etc., and the exact amount is appropriately determined by a doctor.

  The present invention further provides a pharmaceutical composition for treating a patient suffering from a disease for which inhibition of reuptake of norepinephrine is effective. The pharmaceutical composition comprises a therapeutically effective amount of a polygalatenoside represented by the above formula or a pharmaceutically acceptable salt thereof together with a pharmaceutically acceptable carrier and diluent. The polygalatenoside represented by the above formula (1) or a pharmaceutically acceptable salt thereof acts as a norepinephrine reuptake inhibitor by blocking norepinephrine transport. The carrier and diluent used in the pharmaceutical composition can be the same as those described for the antidepressant composition. Further, the administration method (orally and the like), the formulation method, the administration form (tablet and the like), and the dosage are the same as described in the above antidepressant composition.

  The disease to which the composition of the present invention is applied is preferably addictive disorder, withdrawal syndrome, adaptation disorder, age-related learning disorder or mental disorder, anorexia, lethargy, attention deficit disorder, attention deficit hyperactivity Sexual disorder (ADHD), bipolar disorder, or obesity.

Fractionation and purification of [ ^125I ] RTI-55-membrane binding assay of an aqueous extract from the roots of Polygala tenuifolia yielded five novel oligosaccharide derivatives, polygalatenosides AE (compounds 1-5) It was. The structures of these novel oligosaccharide derivatives were analyzed based on spectroscopic evidence. Polygalatenosides A and B (compounds 1 and 2) showed significant inhibitory activity with IC ₅₀ values of 30.0 and 6.04 μM, respectively, in the binding assay according to this example, and as norepinephrine reuptake inhibitors by norepinephrine transport inhibition It became clear that it worked.

[Experimental procedure]
The optical rotation was measured using an optical rotation measuring device (JASCO DIP-370). The UV spectrum was measured using a spectrophotometer (Hitachi UV-3210, manufactured by Hitachi, Ltd.). The IR spectrum was measured using a spectrophotometer (JASCO IR Report-100) by the KBr disc method. HPLC was performed on a Shimadzu LC-10AT _VP system using a Cosmosil® 5C-18-MS-II column (20 × 250 mm and 4.6 × 250 mm, 5 μm). ¹ H, ¹³ C, HMQC, HMBC, and NOESY NMR were performed on a Bruker AMX-400 and Varian-400 Unity Plus NMR spectrometer using tetramethylsilane (TMS) as an internal standard, and all chemical shift units. Is one millionth (ppm, δ). Mass spectra (EI or FAB) were performed on a VG70-250S spectrometer.

[Plant material]
P. Tenifolia roots were purchased at a market in Taipei, Taiwan in May 2003. S. Prof. Kuoh (National Cheng Kung University, Faculty of Life Sciences) confirmed the authenticity. The plant evidence specimen (No. 920021) is deposited in the collection of specimens from the Sakai Industrial Technology Exhibition Center (Taipei, Taiwan).

[Extraction and isolation]
P. Tenifolia (1.25 kg) air-dried roots were powdered and extracted twice with H ₂ O (5 L) for 2 hours under reflux. The H ₂ O extract was applied to a Diaion HP-20 column chromatograph and eluted sequentially with H ₂ O (45 L), 50% MeOH (30 L), and MeOH (25 L). Concentrate the 50% MeOH eluate under reduced pressure to give a pale yellow syrup (44 g) to obtain 6 fractions, CHCl ₃ / MeOH (80:20, 75:25, 70:30, 65:35, and 50: 50) and 100% methanol and chromatographed on a silica gel column (203-400 mesh, E. Merck, 800 g). Recycle high performance liquid chromatography using a mixture of H ₂ O / CH ₃ CN (H ₂ O: CH ₃ CN = 70: 30, flow rate: 10 mL / min; UV 230 nm) using fraction 2 (1.0 g) as a mobile phase. Subjected to preparative HPLC [ODS-5 (20 × 250 mm)], 3 sub-fractions: 2-1 (125.3 mg), retention time 5-13 minutes; 2-2 (121.3 mg), retention time 13-18 minutes, and 2-3 (27.1 mg), retention times 18-23 minutes were obtained. Subfraction 2-2 was separated by HPLC [column: mobile phase H ₂ O-MeOH (80:20) ODS-5 (4.6 × 250 mm); flow rate: 1.0 mL / min; UV: 230 nm] Compound 2 (retention time: 13.3 minutes) (3.1 mg), Compound 5 (retention time: 17.7 minutes) (1.6 mg), Compound 4 (retention time: 19.4 minutes) (3.8 mg) Compound 3 (retention time: 20.9 minutes) (4.6 mg), and Compound 1 (retention time: 22.2 minutes) (33.8 mg) were obtained. The structural formula of each compound is as follows.

[Polygalatenoside A (Compound 1)]
Colorless syrup-like, [α] _D +171 (c 0.01, MeOH); UV (MeOH) λ _max (log ε) 228 (4.12), 273 (3.41), 279 (sh) (3.37) nm IR (KBr) ν _max 3411, 1713, 1634, 1603, 1585, 1285, 1080 cm ⁻¹ ; ¹ H and ¹³ C NMR, see Tables 1 and 2; FABMS m / z 431 ([M + H] ⁺ , 3), 307 (40), 291 (24), 289 (18), 267 (8), 154 (100), 139 (11), 138 (28), 137 (56), 136 (58), 107 (15); HRFABMS m / z 431.1557 [M + 1] ⁺ (calcd for C ₁₉ H ₂₇ O ₁₁ , 431.1553).

[Polygalatenoside B (Compound 2)]
Colorless syrup-like, [α] _D +343.1 (c0.003, MeOH); UV (MeOH) λ _max (log ε) 228 (3.94), 272 (3.69) nm; IR (KBr) ν _max 3415 , 2927, 1713, 1602, 1452, 1280 cm ⁻¹ ; ¹ H and ¹³ C NMR, see Tables 1 and 2; FABMS m / z 431 [M + H] ⁺ , 307, 291, 289, 154, 137, 136, 107; HRFABMS m / z 431.1552 [M + 1] ⁺ (calcd for C ₁₉ H ₂₇ O ₁₁ , 431.1553).

[Polygalatenoside C (Compound 3)]
Colorless syrup-like, [α] _D +256.6 (c0.005, MeOH); UV (MeOH) λ _max (log ε) 229 (3.87), 273 (3.72), 301 (3.55) nm; IR (KBr) ν _max 3402, 1713, 1631, 1602, 1452, 1280 cm ⁻¹ ; ¹ H and ¹³ C NMR, see Tables 1 and 2; FABMS m / z 431 ([M + H] ⁺ , 3.4), 307 ( 33), 291 (21), 289 (15), 267 (8), 155 (27), 154 (100), 139 (11), 138 (29), 137 (57), 136 (61), 107 ( 16); HRFBMS m / z 431.1554 [M + 1] ⁺ (calcd for C ₁₉ H ₂₇ O ₁₁ , 431.1553).

[Polygalatenoside D (Compound 4)]
Colorless syrup-like, [α] _D +103.7 (c0.004, MeOH); UV (MeOH) λ _max (log ε) 216 (3.98), 258 (4.02) nm; IR (KBr) ν _max 3414 , 1708, 1606, 1512, 1464 cm ⁻¹ ; ¹ H and ¹³ C NMR, see Tables 1 and 2; HRABMS m / z 477. 1606 [M + 1] ⁺ (calcd for C ₂₀ H ₂₉ O ₁₃ , 477.611).

[Polygalatenoside E (Compound 5)]
Colorless syrup-like, [α] _D +616.8 (c0.001, MeOH); UV (MeOH) λ _max (log ε) 258 (4.15) nm; IR (KBr) ν _max 3400, 1585, 1505, 1464, 1405cm ^{-1; 1} H and ¹³ CNMR, tables 1 and 2 ^{refer; FABMS m / z 505 ([} M + H] +, 0.5), 503 (2), 459 (3), 371 (3), 369 (3 ), 297 (4), 277 (11), 241 (16), 185 (100), 149 (28), 117 (10), 93 (98), 75 (40); HRFBMS m / z 505.1920 [ M + 1] ⁺ (calcd for C ₂₂ H ₃₃ O ₁₃ : 5055.1923).

[Membrane binding assay]
A membrane derived from canine kidney MDCK cells into which human norepinephrine transporter was stably introduced was used. Whole cell membranes were prepared from transfected cells cultured to confluence in 500 cm ² tissue culture plates. The cells were scraped into a centrifuge tube and precipitated at 900 g, 4 ° C. for 10 minutes. The precipitate was resuspended with a modified Tris-HCl buffer (50 mM Tris-HCl, 100 mM NaCl, 1 μM leupetin, 10 μMPMSF; pH 7.4), and centrifuged at 17000 g, 4 ° C. for 30 minutes. Thereafter, the precipitate was resuspended, homogenized with a glass homogenizer equipped with Teflon pestol, and centrifuged at 17000 g, 4 ° C. for 90 minutes. The precipitate was collected and resuspended in a modified Tris-HCl buffer. The protein concentration was quantified using a BCA protein measurement reagent (Pierce, Rockford). For binding assays, 40 μg aliquots of membrane protein were incubated with 0.2 nM [ ¹²⁵ I] RTI-55 (3β- (4-iodophenyl) -tropan-2β-carboxylic acid methyl ester) at 4 ° C. for 3 hours. Binding was stopped by rapid vacuum filtration using Whatman GF / B soaked with 0.3% polyethyleneimine and three rapid washes with 1 mL cold buffer. Bound radioactivity was measured by gamma spectrometry. Non-specific binding was measured in the presence of 10 μM desipramine and non-specific binding was subtracted from the data in the absence of desipramine to calculate specific binding.

[result]
P. Tenuifolia (1.25 kg) air-dried roots were powdered and extracted with water under reflux. The H ₂ O extract was applied to a Diaion HP-20 column chromatograph, eluting sequentially with H ₂ O, 50% MeOH, and 100% MeOH. The 50% MeOH eluate was chromatographed on a silica gel column with CHCl ₃ / MeOH to obtain 6 fractions. Among those obtained, fraction 2 inhibited [ ¹²⁵ I] RTI-55 against the norepinephrine transporter of MDCK cells in a concentration-dependent manner. The IC ₅₀ value was 4.6 μg / mL. Next, the fraction 2 was subjected to recycle high performance liquid chromatography (preparative HPLC) using a reverse phase ODS column to obtain five new oligosaccharide derivatives polygalatenosides A to E (compounds 1 to 5).

Polygalatenoside A (Compound 1) was isolated as a colorless syrup. The HRFBMS of Compound 1 showed a molecular ion peak protonated at m / z 431.1557 [M + H] ⁺ and was found to be C ₁₉ H ₂₆ O ₁₁ . The ultraviolet spectrum showed maximum absorption indicating the presence of benzoyl residues. In the IR spectrum, it was found that there were bands at 3411 and 1713 cm ⁻¹ and a hydroxyl group and a conjugated ester carbonyl group were present. ^{In the 1} H NMR spectrum, in addition to signals due to galactosyl and polygalitosyl residues, signals for benzoyl groups (δ 7.9, 2H d; 7.61, 1H, t; 7.47, 2H, d) appeared. (See Table 1). The ¹³ C NMR signal (see Table 2) with benzoyl, galactosyl, and polygalitosyl moieties also revealed that Compound 1 is a polygalitosyl galactoside of benzoic acid. All proton and carbon NMR signals were determined by ¹ H- ¹ H COSY, HMQC, and HMBC NMR experiments. Compared to polygalitol (the following formula (3)), C-2 of the polygalitosyl unit is shifted to a low magnetic field of 4 ppm, and C-1 of the galactosyl unit is shifted to δ _C 95.7 on the low magnetic field side. It was revealed that there is a glycoside bond in Compound 1 as polygalitosyl (2 → 1) -α-galactoside.

This is also supported by the ³ J correlation between H-1 (δ _H 5.00) of the galactosyl unit and C-2 (δ _C 73.3) of the polygalitol moiety in the HMBC experiment (Fig. 1). In the NMR spectrum, the methylene proton signal of the galactosyl residue is low field shifted to δ _H 4.49 and 4.39, and the C-6 carbon of the galactosyl residue is low field to δ _C 64.3. From the shift shift, it is determined that the C-6 position is a benzoyl group bonding position. This was confirmed by _H-6 (δ H 4.49 and 4.39) and ³ J coupling constant between the ester carbonyl carbon of the benzoyl residues _{(δ C 168.2) (correlation)} . From the analysis of the above data, polygalatenoside A (compound 1) was found to be 6-O-benzoyl-polygalitosyl (2 → 1) -α-galactose (6-O-benzoyl-polygalitosyl- (2 → 1) -α-galactose). It was judged that.

Polygalatenoside B (compound 2) and polygalatenoside C (compound 3) were isolated as a colorless syrup. HRABMS of compounds 2 and 3 showed molecular ion peaks protonated at m / z 431.1552 and 431.1554 [M + H] ⁺ , respectively, and were found to be C ₁₉ H ₂₆ O ₁₁ as in compound 1. In the ultraviolet spectrum, the maximum absorption corresponding to the benzoyl residue was shown. In the IR spectrum, it was found that there were bands at 3415 and 1713 cm ⁻¹ , and hydroxyl groups and conjugated ester carbonyl groups were present. In the NMR spectra of polygalatenosides B and C, the presence of a benzoyl residue as the ester moiety and the presence of galactosyl and polygalitosyl residues as the sugar moiety as in compound 1 was revealed. After allocating all the proton signals from the ¹ H- ¹ H COZY and HMQC spectra, the substitution positions of these residues were determined by NOE (see FIG. 2) and HMBC (see Fig. 1). ^{In the 13} J NMR correlation and the ³ J correlation between H-1 (galactosyl) and C-2 (polygalitosyl) in the HMBC spectra of compounds 2 and 3, the C-2 of the polygalitosyl moiety and the C- of the galactosyl moiety From the fact that 1 shifts in a low magnetic field, it was revealed that a sugar residue exists as polygalitosyl (2 → 1) -α-galactoside as in Compound 1. However, compounds 2 and 3 were found to differ in the ester binding site. In compound 2, since the signals of H-3 and C-3 of the galactosyl unit are shifted in a low magnetic field to δ _H 5.29 and δ _C 75.3, benzoyl is present at the C-3 position of the galactosyl residue. The group was found to exist. In compound 3, the H-4 and C-4 signals of the galactosyl unit are shifted in a low magnetic field to δ _H 5.59 and δ _C 73.5, respectively, and the benzoyl group is bound at the position of galactosyl C-4. It became clear that. This is also supported in the HMBC spectrum by the ³ J correlation between the galactosyl unit H-4 (δ _H 5.59) and the carbonyl carbon C-7 of the benzoyl group (δ _C 167.9). . Thus, structurally, compound 2 is 3-O-benzoyl-polygalactosyl- (2 → 1) -α-galactose (3-O-benzoyl-polygalactosyl- (2 → 1) -α-galactose) and compound 3 is 4 It was found to be -O-benzoyl-polygalactosyl- (2 → 1) -α-galactose (4-O-benzoyl-polygalactosyl- (2 → 1) -α-galactose).

Polygalatenoside D (compound 4) was obtained in the form of a colorless syrup. HRABMS showed a molecular ion peak protonated at m / z 477.1606 and was found to be C ₂₀ H ₂₉ O ₁₃ . The ultraviolet spectrum of Compound 4 showed absorption at 216 and 258 nm. In the IR spectrum, it was found that there were bands at 3414 and 1708 cm ⁻¹ , and hydroxyl groups and conjugated ester carbonyl groups were present. ¹ HNMR spectra for A ₂ B ₂ -type aromatic protons (δ 8.06 and 7.01 for each 2H) and methoxy groups (δ 3.86, 3H, s) in addition to the signal for the sucrose moiety. A signal was shown. The ¹³ C NMR spectrum of Compound 4 also showed a signal due to the p-methoxybenzoyl group and sucrose residue (see Table 2). All assignments of ¹ H and ¹³ C NMR signals were confirmed by COSY, HMQC, and HMBC experiments. Since the oxymethine proton and carbon (H-3 and C-3) of the fructosyl moiety are low field shifted to δ _H 5.56 and δ _C 80.6 respectively, the p-methoxybenzoyl moiety is It was shown to be located at C-3 ′. This is supported by the HMBC spectrum. This is because the fructosyl residue H-3 (δ _H 5.56) correlates to the ester carbonyl carbon of the p-methoxybenzoyl moiety at δ _C 167.7. Therefore, the structure of the compound 4 is presumed to be 3′-Op-methoxybenzoyl-sucrose (3′-Op-methoxybenzoyl-sucrose).

Polygalatenoside E (compound 5) is isolated in a colorless syrup-like form and is presumed to have an elemental composition of C ₂₂ H ₃₂ O ₁₃ from HRFABMS ([M + H] ⁺ m / z 505.1920). IR absorption bands at 3400, 1585, 1505, 1464 cm ⁻¹ indicate the presence of hydroxyl and aromatic moieties. The NMR data of Compound 5 showed a cis-sinapylic alcohol partial signal. The cis-sinapyrcohol partial signal is glucosyl and δ _C at ¹³ C NMR spectrum of Compound 5 at δ _C 103.2, 79.2, 79.2, 71.8, 78.5 and 63.1 In addition to the signal of apiosyl at 110.9, 78.6, 81.4, 76.1 and 66.9, two equivalent aromatic protons at δ _H 6.55 (2H, s) , Two methoxy groups at δ _H 3.83 (6H, s), δ _H 6.48 (1H, d, J = 11.6 Hz) and 5.80 (1H, dt, J = 11.6, 6) 2 cis-olefin protons at 4 Hz) as well as oxymethylene protons at δ _H 4.34 (2H, dd, J = 6.4, 1.6 Hz). The above data and caropanaxin D [Kazuko, S .; Shuichi, S .; Yoshiteru I .; Junzo, S .; Chem. Pharm. Bull. 1991, 39, 865-870), the sugar moiety is presumed to be a β-apiosyl- (1 → 2) -β-glucoside (β-apiosyl- (1 → 2) -β-glucoside) moiety. Is done. This is due to the low field shift of glucosyl C-2 to δ _C 79.2 and the H-2 proton (δ _H 3.69) of the glucosyl unit and C-1 (δ _C 110. 9) supported by the ³ J correlation (see FIG. 1). In addition, based on what was observed in the HMBC correlation between glucosyl H-1 (δ _H 5.10) and cis-sinapyr C-4 (δ _C 135.5), sinapyl alcohol was converted to glucosyl C-1. To position. From the above data, compound 5 is sinapyl alcohol 4-O-β-apiosyl- (1 → 2) -β-glucose (4-O-β-apiosyl- (1 → 2) -β-glucose). Became clear.

Whether compounds 1-5 inhibit the binding of isotope-labeled RTI-55 to norepinephrine transporter protein was tested in vitro [Galli, A. et al. Defelice L .; J. et al. Duke, B .; J. et al. Moore, K .; R. Blakely, R .; D. J. et al. Exp. Biol. 1995, 198, 2197-2212]. In the membrane binding assay, polygalatenoside A (Compound 1) and B (Compound 2) showed significant inhibitory activity with IC ₅₀ values of 30.0 and 6.04 μM, respectively. Decipramine, a tricyclic antidepressant, inhibited [ ¹²⁵ I] RTI-55 binding to NET with an IC ₅₀ value of 0.93 nM. From the above results, it was shown that the compound of the present invention acts as a norepinephrine reuptake inhibitor by specifically inhibiting the binding to NET.

It is a figure which shows HMBC (Heteronuclear Multiple Bond Correlation) correlation of the compounds 1-5 of this invention refine | purified from the aqueous solution extract from the root of Polygala tenufolia. FIG. 2 is a diagram showing a NOESY (Nuclear Overhauser Effect Spectroscopy) correlation of Compounds 2 and 3 of the present invention purified from an aqueous extract from roots of Polygala tenuifolia.

Claims (5)

The following chemical formula (1):
In the formula, R, R ′ and R ″ are each independently a hydrogen atom or R ₁ (in this case, R, R ′ and R ″ are not all hydrogen atoms), and R ₁ is Chemical formula (2);
In the formula, R ₂ represents one selected from the group consisting of a hydrogen atom, a C1-6 alkyl group, a C1-6 alkoxy group, and a halogen atom, or a pharmaceutically acceptable salt thereof. The compound according to claim 1, wherein R ₂ is a hydrogen atom, or a pharmaceutically acceptable salt thereof. The compound or a pharmaceutically acceptable salt thereof according to claim 2, wherein any one of R, R 'and R "is R ₁ and the other two are hydrogen atoms.   Norepinephrine reuptake comprising the compound of any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof as a norepinephrine reuptake inhibitor by blocking norepinephrine transport together with a pharmaceutically acceptable carrier and diluent. A pharmaceutical composition for diseases effective for inhibition. The disease is addictive disorder, withdrawal syndrome, adaptation disorder, age-related learning disorder or mental disorder, anorexia, lethargy, attention deficit disorder, attention deficit hyperactivity disorder (ADHD), bipolar disorder, or obesity The pharmaceutical composition according to claim 4 .